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Early detection and early slaughter through quarantine are essential to prevent the spread of the African swine fever virus (ASFV). Pooled oral fluids testing is a practical approach for pathogen monitoring, but compared to blood, the virus concentration is lower, increasing false negatives.
Objective: We developed a highly sensitive method for ASFV detection in pig oral fluids using a pretreatment-based concentration protocol.
Methods: We collected oral fluids from sub-clinical raised pigs in northern Vietnam and conducted a field evaluation using the developed method. A spike test showed up to 100-fold greater sensitivity than a reference method. For performance evaluation, 68 pooled oral fluid samples were collected, of which 63 originated from raised pigs in northern Vietnam and 5 were obtained from healthy pigs in Japan as negative controls.
Results: Using real-time PCR, 9/68 (13.2%) were positive by the reference method, while 23/68 (33.8%) were positive by the developed method. Real-time LAMP detected 1/68 (1.5%) and 6/68 (8.8%) respectively. The developed method therefore improved the diagnostic performance and enabled early diagnosis prior to disease onset.
Conclusion: These results suggest enhanced sensitivity and feasibility for early ASF diagnosis, potentially contributing to more effective outbreak control. The developed method offers a simple and sensitive tool for rapid ASFV screening in both affected and at-risk farms.
Mai TN, Tran THG, Dong VH et al. A highly sensitive method for detecting African swine fever virus in oral fluids from naturally infected pigs in Northern Vietnam. Sci Rep 15, 27855 (2025). https://doi.org/10.1038/s41598-025-12139-8
